title: Engineering an enantioselective amine oxidase for the synthesis of pharmaceutical building blocks and alkaloid natural products.
abstract: The development of cost-effective and sustainable catalytic methods for the production of enantiomerically pure chiral amines is a key challenge facing the pharmaceutical and fine chemical industries. This challenge is highlighted by the estimate that 40-45% of drug candidates contain a chiral amine, fueling a demand for broadly applicable synthetic methods that deliver target structures in high yield and enantiomeric excess. Herein we describe the development and application of a "toolbox" of monoamine oxidase variants from Aspergillus niger (MAO-N) which display remarkable substrate scope and tolerance for sterically demanding motifs, including a new variant, which exhibits high activity and enantioselectivity toward substrates containing the aminodiphenylmethane (benzhydrylamine) template. By combining rational structure-guided engineering with high-throughput screening, it has been possible to expand the substrate scope of MAO-N to accommodate amine substrates containing bulky aryl substituents. These engineered MAO-N biocatalysts have been applied in deracemization reactions for the efficient asymmetric synthesis of the generic active pharmaceutical ingredients Solifenacin and Levocetirizine as well as the natural products (R)-coniine, (R)-eleagnine, and (R)-leptaflorine. We also report a novel MAO-N mediated asymmetric oxidative Pictet-Spengler approach to the synthesis of (R)-harmicine.
authors.affiliation: School of Chemistry, Manchester Institute of Biotechnology, University of Manchester, 131 Princess Street, Manchester M1 7DN, United Kingdom.
authors.forename: Diego
authors.initials: D
authors.lastname: Ghislieri
authors.forename: Anthony P
authors.initials: AP
authors.lastname: Green
authors.forename: Marta
authors.initials: M
authors.lastname: Pontini
authors.forename: Simon C
authors.initials: SC
authors.lastname: Willies
authors.forename: Ian
authors.initials: I
authors.lastname: Rowles
authors.forename: Annika
authors.initials: A
authors.lastname: Frank
authors.forename: Gideon
authors.initials: G
authors.lastname: Grogan
authors.forename: Nicholas J
authors.initials: NJ
authors.lastname: Turner
mesh_terms: D000470:Alkaloids / Q000737:chemistry / Q000378:metabolism*; D001234:Aspergillus niger / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D001559:Benzhydryl Compounds / Q000737:chemistry / Q000378:metabolism; D001688:Biological Products / Q000737:chemistry / Q000378:metabolism; D026121:Indole Alkaloids / Q000737:chemistry / Q000378:metabolism; D008958:Models, Molecular; D008995:Monoamine Oxidase / Q000235:genetics* / Q000378:metabolism; D010084:Oxidation-Reduction; D015202:Protein Engineering / Q000379:methods*; D013237:Stereoisomerism
chemical_list: D000470:Alkaloids; D001559:Benzhydryl Compounds; D001688:Biological Products; D026121:Indole Alkaloids; C544842:harmicine; C004531:benzhydrylamine; D008995:Monoamine Oxidase